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Objective To investigate the inhibitory effect of bone marrow mesenchymal stem cells ( BMMSC) on the inflammatory response of microglial cells .Methods The samples were divided into four groups .Group I:microglial (BV-2) cells were cultured in DMEM (high glucose).Group II: BV-2 cells were cultured in DMEM containing lipopo-lysaccharide (LPS).Group III:BV-2cells and BMMSCs were co-cultured in DMEM.Group IV: BMMSCs were cultured in DMEM containing LPS .The growth state and ultrastructure of BV-2 cells were observed and the changes of TNF-αex-pression were detected .Results Different cell densities of BV-2 cells were observed under the optical microscope in an or-der from high level to low level:group I >group III >group II.The expressions of TNF-αwere:group Ⅱ >groupⅢ>group Ⅰ.Ultrastructural observation of BV-2 cells showed that there were a large number of swollen mitochondria and endoplasmic reticulum , some of them showed vacuolization .No BV-2 cells with multiple hucleoli were found in the group II indicating the absence of active cell growth .At the same time, cytolysis was observed only in the group II .The growth of BV-2 cells in the group III was better than that in the group II .Conclusions BMMSCs can inhibit inflammatory response of microglial cells, therefore, play a neuroprotective role.
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BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)have the properties involving high proliferation capability,widely distribution,functional tissue repair after injury,as well as immune modulation,by which bring us extensive therapeutic possibilities.There are plenty of methods for isolation of BMSCs,yet,BMSCs exhibit discrepancies in varied growth stage and culture conditions.Up to now,there has been no agreement about the identification methods for cultured BMSCs.OBJECTIVE:To review the isolation methods and biological characteristics of BMSCs,and to compare the differential expression of BMSCs between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.METHODS:A computer-based online search was performed using key words of "bone marrow mesenchymal stem cells,isolation,culture,induce,marker,and characterization" to find documents published in the database of CNKI (http://dlib.cnki.net/kns50/)or Pubmed(http://www.ncbi.nlm.nih.gov/PubMed)from January 2003 to June 2009.The languages were limited Chinese and English.A total of 237 literatures were searched by the computer.RESULTS AND CONCLUSION:The positive rates of CD44 and CD34 of BMSCs isolated by the whole bone marrow culture were smaller than that of the density gradient centrifugation.However,BMSCs isolated by the whole bone marrow culture were superior to those isolated by the density gradient centrifugation in cell viability,proliferation rate,confluence time,as well as generation time.Other methods for BMSCs isolation had drawbacks of large cost and high requirement of experimental equipments.Following conditions were used to identify BMSCs:cell adherence,cell surface molecule labeling,strong self-proliferation ability,as well as potentials multi-directional differentiation.BMSCs exhibit differential expression between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.
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@#Objective To investigate induction and repair of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) sponge to endogenous neural stem cells after acute spinal cord half cut-off in rats.MethodsThe differences of proliferation and differentiation of endogenous neural stem cells were compared between injured group and intervention treatment group.ResultsThere are remarkable differences between the injured group and intervention treatment group.ConclusionbFGF and EGF sponge can enhance the proliferation and differentiation of the neural stem cells in the test animal.
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@#Objective To investigate the curative effect of neural stem cells (NSCs) transplantation on sequela after traumatic intracranial hematoma. Methods 20 patients with sequela after traumatic intracranial hematoma were treated with NSCs transplantation. Cells were engrafted into subarachnoid cavity via lumbar puncture. They were assessed with Functional Independence Measure (FIM) before and half a year after the transplantation. Results The FIM scores were significantly increased after the transplantation (P<0.01).Conclusion NSCs transplantation could promote functional recovery and improve the living quality of patients with sequela after traumatic intracranial hematoma in the aspects of self-care, sphincter control, mobility, locomotion, communication and social adjustment/cooperation.
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Objective To explore the possibility of repairing injured facial nerve with tissue engineering technology and neural stem cells(NSCs).The complex consisted of NSCs,scaffold and NT-3.NSCs were immature cells with the potential of self-renewal and multiple differentiation to neurons and glial cells.The scaffold with porous surface was made of hyaluronic acid and collagen(HA-Col gel) which degenerate in vivo after transplantation.NT-3 is the signal to promote neurons survival in vitro.Methods NSCs of S-D rat were co-cultured with scaffold and NT-3 in vitro.The two stumps of disconnected facial nerve of rabbit were re-connected with the complex.Electrophysiology and morphology tests were used to examine functional and morphological changes.Results Result] NSCs adhered to the HA-Col gel and survived.Injured facial nerve fixed by NSCs-HA-Col gel-NT-3 complex showed significant improvement in function and anatomical structure.Conclusion Combinative implant of NSCs,HA-Col gel and NT-3 may promote the regeneration of injured facial nerve.
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@#Neurotrophins promote and modulate the repair and regeneration of central nervous system (CNS) on the levels of synapses, neurites and neural cells, and even of the auxiliary structures of CNS. As to immune molecules, recent studies denied the classical doctrine that CNS is an immune privilege organ. In the last decade, it is found that immune responses can be beneficial for CNS repair. What are more, some macromolecules that were previously thought to be the members of the family of immune system play essential roles in the development and regeneration of the CNS. Therefore, the authors postulate that there are crosstalks between the neurotrophins and the immune molecules in the CNS.
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Objective To develop a stable model of focal cerebral infarction in rat to study the curative effect of neural stem cells transplantation.Methods Thirty-seven rats were selected which were divided into two groups in random, experimental group and control group. The focal infarction model was developed by the ligation of the left middle cerebral artery followed by the ligation of the ipsilateral common carotid artery and the temporary clip occlusion of the contralateral common carotid artery for 1.5 h. The operation adopted minimally invasive craniotomy though temporal bone. The model was evaluated by examining the neurologic deficits, ink perfusion, TTC staining and Magnetic Resonance imaging.Results All the rats were in good condition after the operation, the mortality rate was 6.25% after 4 weeks. Ink perfusion and TTC staining confirmed that the ischemia was confined to the cortex. The areas of infarction measured 83.52 mm3 by Magnetic Resonance imaging after 4 weeks.Conclusion A stable focal cerebral infarction model can be achieved by minimally invasive craniotomy. It is superior for its homogeneity of infarction volume and site, and its low mortality. It can be used for the study of transplantation of neural stem cells.
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@# ObjectiveTo explore environmental conditions under which bone marrow stromal cells could be induced into osteogenic phenotype.MethodsRat bone marrow stromal cells were isolated and proliferated in vitro, and the 3rd passage was divided into the group A (control group), group B (cells cultured in the medium containing dexamethasone, β-glycerol disodium phosphate salt hydrate, vitamin C and active form of vitamin D3), and group C (on the bases of group B, the cells were cultured additionally with fracture hematoma extract). On the post-induction day 5, 8, and 11, the morphological changes were observed and the osteogenic markers such as alkaline phosphotase (ALP), collagen type Ⅰ (Col Ⅰ) and osteocalcin (OCN) were assayed with immunohistochemical staining, the calcification was manifested with von Kossa staining.ResultsIn the group A, no evident osteogenic effects had been observed. In the group B, on 5th day post-induction, some bone marrow stromal cells underwent a morphological change, and mild expression of ALP and Col Ⅰ was observed but with no calcification effect. On 8th day post-induction, the ratio of morphologically changed cells increased, and the expression of ALP and Col Ⅰ increased still with no evident calcification. On 11th day post-induction, anti-OCN staining was positive and the calcium nodes were showed by von Kossa staining. The phenotype changes in the group C were similar to group B, but were more evident.ConclusionBone marrow stromal cells can be induced into osteogenic phenotype in vitro with small molecular inducers. Fracture hematoma extract can enhance this effect thus might be used as an addictive in osteogeneration.
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BACKGROUND: Both animal experiments and clinical practice have confirmed that mild or moderate hypothermia is effective in reducing secondary brain injury, but its effect on homeostasis is not very clear.OBJECTIVE: To investigate the effect of a combined therapy of mild hypothermia and hibernation on the homeostasis of patients with severe brain injury.DESIGN: A randomized controlled study.SETTING: Neurosurgical Institute of Beijing; Neurosurgical Department of the First Clinical Medical College Affiliated to Harbin Medical University;and Neurological Department of the Second Clinical Medical College Affiliated to Harbin Medical University.PARTICIPANTS: The study was conducted at the Department of Neurosurgery, the First Affiliated Hospital of Harbin Medical University, from June to December 2002. Totally 24 patients (aged 35-60 years) with severe cerebral hemorrhage or brain injury were randomly divided into combined therapy group and normothermia group. Their Glasgow Coma Scale scores ranged from 3 to 8. The subjects signed the informed consent.METHODS: Within 10 hours of their injury, patients in hypothermia and hibernation combination group were given half dosage of No. 1 hibernation cocktail (chlorpromazine 25 mg, pethidine hydrochloride 50 mg, and promethazine 25 mg), and were cooled by cooling blankets to make their body temperature dropped to 32-34 ℃ (rectal temperature). Their temperature was kept within this range for 5 days, at 35 ℃ for 24 hours, and then was slowly increased to their normal level. The body temperature of patients in normothermia group was maintained at 37-38 ℃. The mean arterial pressure and heart rate of all patients were measured continuously by HP monitor. On the 3"d and 7th days of hospitalization, intracranial pressure and creatine phosphate kinase were measured via lumbar puncture.Femoral artery puncture was performed to check the partial pressure of arterial O2 and CO2. Platelets count and blood electrolytes K+ and Na+ concentration of each patient were measured, too. On the 7th day Glasgow Outcome Scale scores of each patient and mortality of each group were recorded.activity of creatine phosphokinase, platelets count, blood K+ and Na+ conand CO2 of patients in combined therapy group on the 3rd and 7th days of hospitalization.intracranial pressure, creatine phosphokinase and platelets count: The decreased values of intracranial pressure, creatine phosphokinase and platelet number in combined therapy group were all significantly higher than those in normothermia group [(104.09±54.90), (58.75±25.33) mm H2O; (26.95±19.22), (10.17±7.18) μkat/L; (89.82±46.36)×109/L, (48.83±44.59)×109/L,the mean arterial pressure, blood electrolytes, and partial pressure of artewas significantly lower than that of normothermia group (25.0%, 66.6%,P ＜0.05).CONCLUSION: This combined therapy of hypothermia and hibernation can effectively decrease intracranial pressure and creatine phosphokinase,but has no significant effects on the mean arterial pressure, blood electrolytes concentration, and partial pressure of arterial O2 and CO2. It has the risk of disturbing the patients' hematopoiesis.
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<p><b>OBJECTIVE</b>To explore the factors which induce differentiation of embryonic neural stem cells.</p><p><b>METHODS</b>Rat embryonic neural stem cells were co-cultured with newborn rat Schwann cells in serum-free medium. The phenotype and specific-markers including tubulin-beta, glial fibrillary acidic protein (GFAP) and galactorcerebroside (GalC), were demonstrated by phase contrast microscopy and double immunofluorescence staining.</p><p><b>RESULTS</b>Overall, 80% +/- 5% of neural stem cells protruded several elongated processes and expressed tubulin-beta antigen at high levels, while 20 +/- 3% of them protruded several short processes and were GalC or GFAP positive.</p><p><b>CONCLUSION</b>The factors secreted by Schwann cells could induce rat embryonic neural stem cell to differentiate.</p>
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Animals , Rats , Cell Differentiation , Coculture Techniques , Embryo, Mammalian , Cell Biology , Fluorescent Antibody Technique , Microscopy, Phase-Contrast , Nerve Growth Factors , Physiology , Neurons , Cell Biology , Phenotype , Rats, Sprague-Dawley , Schwann Cells , Physiology , Stem Cells , Cell BiologyABSTRACT
@#ObjectiveTo investigate whether intracerebral transplantation of heterogenous neural stem cells(NSCs) may cause immunal rejection or not.MethodsNSCs derived from the brain of embryonic 16-day Wistar rat were cultured in vitro. Cerebral ischemic SD rat models were made by middle cerebral artery occlusion and NSCs were transplanted into the ipsilateral or contralateral caudate nucleus 4 days later. Rats were sacrificed at different time points after transplantation, the reaction and proliferation of mononuclear phagocytic system(MPS) and splenic follicles as well as the survival and migration of NSCs were checked.ResultsNumerous NSCs survived and migrated toward the lesion in vivo. Comparing with control group that did not receive NSCs transplantation, there was no significant infiltration of mononuclear phagocytes or proliferation of splenic follicles in NSCs transplanted rats.ConclusionsIntracerebral transplantation of heterogenous neural stem cells did not cause remarkable immunal rejection.